CN103476923A - A method for promoting the synthesis of collagen and proteoglycan in chondrocytes - Google Patents

A method for promoting the synthesis of collagen and proteoglycan in chondrocytes Download PDF

Info

Publication number
CN103476923A
CN103476923A CN2011800495672A CN201180049567A CN103476923A CN 103476923 A CN103476923 A CN 103476923A CN 2011800495672 A CN2011800495672 A CN 2011800495672A CN 201180049567 A CN201180049567 A CN 201180049567A CN 103476923 A CN103476923 A CN 103476923A
Authority
CN
China
Prior art keywords
intervertebral disc
vaccinia virus
treatment
inflammation
tissue extract
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN2011800495672A
Other languages
Chinese (zh)
Other versions
CN103476923B (en
Inventor
舛田浩一
内木充
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nippon Zoki Pharmaceutical Co Ltd
University of California
Original Assignee
Nippon Zoki Pharmaceutical Co Ltd
University of California
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nippon Zoki Pharmaceutical Co Ltd, University of California filed Critical Nippon Zoki Pharmaceutical Co Ltd
Publication of CN103476923A publication Critical patent/CN103476923A/en
Application granted granted Critical
Publication of CN103476923B publication Critical patent/CN103476923B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/32—Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5082—Supracellular entities, e.g. tissue, organisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/01—DNA viruses
    • G01N2333/065—Poxviridae, e.g. avipoxvirus
    • G01N2333/07—Vaccinia virus; Variola virus

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Cell Biology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Rheumatology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Toxicology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dermatology (AREA)
  • General Engineering & Computer Science (AREA)
  • Tropical Medicine & Parasitology (AREA)

Abstract

The synthesis of collagen and proteoglycan in chondrocytes, such as intervertebral disc cells, articular chondrocytes and meniscal cells is promoted by administration of an extract from inflamed tissue inoculated with vaccinia virus.

Description

The promotion method that collagen protein in the chondrocyte and proteoglycan are synthetic
Technical field
The present invention relates to inoculate the new medicine use of the inflammation tissue extract of vaccinia virus, particularly the chondrocyte is as collagen protein and the synthetic promotion method of proteoglycan in intervertebral disk (intervertebral disc cell) cell, articular chondrocytes (articular chondrocyte) and meniscus cell (meniscus cell).
Background technology
The kit that the trade(brand)name " Neurotropin (Neurotropin) " of usining is sold containing the non-protein extract of the inflammation skin of the rabbit that vaccinia virus is arranged from inoculation as effective constituent.Be widely used in the treatment neurogenic pain at Japanese Neurotropin (NTP), comprised postherpetic neuralgia and pain in the back.In animal model, NTP has demonstrated anti-nociception (anti-nociceptive) effect that the activation due to Descending monoamine energy pain inhibition system causes.NTP also demonstrates by suppressing the activation of kallikrein-kassinin kinin (kallikrein-kinin) cascade, suppresses thus bradykinin and comes the inhibition of pain path with external formation in vivo.
Reported recently the inhibition of NTP to the mrna expression of the tumor necrosis factor alpha (TNF-α) of human disc cell and cyclooxygenase-2 (COX-2).Yet, do not know whether NTP has the synthetic effect of extracellular matrix (ECM) to intervertebral disc cells.
Summary of the invention
Be used for promoting the chondrocyte as the collagen protein of intervertebral disc cells (ox nucleus pulposus cell (bovine nucleus pulposus cell) or annulus fibrosis cells (annulus fibrosus cell)), articular chondrocytes and meniscus cell and synthesizing of proteoglycan the inflammation tissue extract of inoculation vaccinia virus.Aspect of the present invention, proteoglycan in the chondrocyte who promotes the patient and/or the method for collagen protein synthesis are provided, described method comprises to the inflammation tissue extract of the inoculation vaccinia virus of patient's administration pharmacology significant quantity of this type for the treatment of of needs.In another aspect of this invention, provide the method for the extracellular matrix of regeneration of cartilage cell, described method comprises to patient's administration of this type for the treatment of of needs the extract of the inflammation separate tissue of vaccinia virus from inoculation.
The accompanying drawing explanation
Further illustrate the present invention by accompanying drawing, wherein:
Fig. 1 is the experimental result of inflammation tissue extract synthetic activity to proteoglycan in the intervertebral disc cells as annulus fibrosis cells (AF) of inoculation vaccinia virus.
Fig. 2 is the experimental result of inflammation tissue extract synthetic activity to proteoglycan in the intervertebral disc cells as ox nucleus pulposus cell (NP) of inoculation vaccinia virus.
The experimental result of the inflammation tissue extract that Fig. 3 is the inoculation vaccinia virus to the activity of collagen protein synthesis in the intervertebral disc cells as annulus fibrosis cells (AF).
The experimental result of the inflammation tissue extract that Fig. 4 is the inoculation vaccinia virus to the activity of collagen protein synthesis in the intervertebral disc cells as ox nucleus pulposus cell (NP).
Embodiment
The present invention so that be used in normal oxygen (normoxic) and hypoxemia (hypoxic) state under the ox nucleus pulposus cell (NP) cultivated in alginates bead (bead) and annulus fibrosis cells (AF) to estimate NTP be basic on the result of the impact of proteoglycan (PG) and collagen protein synthesis.
Inflammation tissue extract about inoculation vaccinia virus of the present invention, as the existence reported before for the physiologically active substance produced in the inflammation tissue that vaccinia virus is arranged in inoculation, from pathological tissues, the various reports of the method for this material of extraction and pharmacologic activity etc. are (referring to [0008] section of WO2009/028605, EP2191836, Japan patent applicant announce JP-A-53-101515, JP-A-55-87724, JP-A-1-265028, JP-A-1-319422, JP-A-2-28119, JP-A-7-97336, JP-A-8-291077, JP-A-10-194978, JP-A-11-80005, JP-A-11-139977, JP-A-2000-336034, JP-A-2000-16942 and JP-A-2004-300146, and international publication number WO2004/039383).
In addition, from inoculation, there is the preparation of extract of inflammation skin of the rabbit of vaccinia virus to be obtained commercially as medicine, and can be used for the present invention.As " Drugs in Japan; Ethical Drugs " (2010, by the medical information center of Japan (Japan Pharmaceutical Information Center) editor distribution) the 2978-2980 page put down in writing, said preparation is to comprise the medicament that extracts the non-protein-active material of separation from inoculation has the inflammation skin histology of rabbit of vaccinia virus.Known said preparation effectively resists pain in the back, cervical rib syndrome, symptomatic neuralgia, scapulohumeral periarthritis (periarthritis scapulohumeralis), osteoarthritis, follow the itch of tetter (eczema, dermatitis, urticaria), allergic rhinitis, the sequela of subacute-myelo-optico-neuropathy (subacute myelo-optico-neuropathy) is as cold, paresthesia and pain, and postherpetic neuralgia etc.The approval said preparation is as the prescription drugs of the form of the form of subcutaneous injection, intramuscular injection and intravenous injection product and tablet, and is obtained commercially.
The non-protein biology function Auto-regulator of inflammation tissue extract for having the inflammation tissue of vaccinia virus to extract from inoculation as mentioned above for inoculation vaccinia virus of the present invention, and in " Drugs in Japan; Ethical Drugs ", list from inoculation, have the extracting solution preparation of inflammation skin of the rabbit of vaccinia virus to be approved for medicine, and be obtained commercially.In addition, that in above-mentioned patent documentation, puts down in writing has the various extracts of the inflammation tissue of vaccinia virus to can be used as material of the present invention from inoculation, and their production method and suitable dosage etc. also provide in the literature.
Method obtains the inflammation tissue extract of inoculation vaccinia virus of the present invention as follows: inoculation is had to the inflammation tissue crushing of vaccinia virus; Add the extraction solvent and remove fragment of tissue; Then carry out the deproteinated processing; By the deproteinated solution absorbs to sorbent material; Follow wash-out effective constituent.
For example according to following method, produce the inflammation tissue extract of inoculation vaccinia virus.
(a) collect the inflammation skin histology that inoculation has the rabbit of vaccinia virus or mouse etc., and the inflammation tissue is crushed.Add to extract solvent as water, phenol water (phenolated water), physiological saline or be added with the glycerol liquor (phenol-added glycerin water) of phenol to the tissue of crushing.Then, filter or centrifugal mixture, thereby obtain extracting solution (filtrate or supernatant liquor).
(b) pH of extracting solution is adjusted into to acid state, and heating liquid carries out the deproteinated processing.Then, deproteinated solution is adjusted into to alkaline state, heating, then filtration or centrifugal.
(c) gained filtrate or supernatant liquor are made to acidity, and be adsorbed onto sorbent material as on gac or kaolin.
(d) add and extract solvent as water to sorbent material, pH is adjusted into to alkaline state, the wash-out absorbed component, thus obtain the inflammation tissue extract of inoculating vaccinia virus.Subsequently, according to expectation, thereby can make elutriant vapourisation under reduced pressure to drying or lyophilize provide dry-matter.
About the animal for by inoculation vaccinia virus acquisition inflammation tissue, the various animals that can use infection that vaccinia virus is arranged are as rabbit, ox, horse, sheep, goat, monkey, rat or mouse, and preferred inflammation is organized as the inflammation skin histology of rabbit.
The inflammation tissue of collecting and crush, and add the extraction solvent of 1-5 volume, thus preparation emulsification suspension.About extracting solvent, can use distilled water, physiological saline and weak acid to the weakly alkaline damping fluid etc.Can suitably add stablizer as glycerine, antibiotic/sanitas is as phenol, and salt is as sodium-chlor, Repone K or magnesium chloride.Now, can be by by processing as freeze thawing, ultrasonic wave, cytolemma lytic enzyme or tensio-active agent destroy cell tissue, thus promote extraction.
Gained emulsification extracting solution is filtered or centrifugal etc., thereby remove fragment of tissue, then carry out the deproteinated processing.Deproteinated operation can be undertaken by known method usually, but application examples as thermal treatment, the processing by protein denaturant as acid, alkali, urea and guanidine, the processing by organic solvent as acetone, isoelectric precipitation and saltouing.Then, general method by removing insolubles for example, as (used filter paper, Mierocrystalline cellulose or nitrocotton), the filtration of glass filter, celite (Celite) or Cai Shi (Seitz) filter etc., ultrafiltration and centrifugal, remove the insoluble protein of separating out.
To comprise the extracting solution acidifying of the effective constituent obtained in this mode, preferably with sour example hydrochloric acid, sulfuric acid or Hydrogen bromide, pH will be adjusted to 3.5-5.5, then be adsorbed onto on sorbent material.The example of available sorbent material comprises gac and kaolin.Can under agitation sorbent material be added in extracting solution, or extracting solution can be passed and is filled with the post of sorbent material, thereby effective constituent is adsorbed onto on sorbent material.When adding in extracting solution by sorbent material, remove solution by filter or centrifugal etc., thereby obtain the sorbent material that wherein absorption has effective constituent.
For from sorbent material wash-out (desorption) effective constituent, add eluting solvent to sorbent material, thereby at room temperature or by suitable heating or by stirring carry out wash-out, and remove sorbent material by general method as filter or centrifugal etc.About the eluting solvent that will use, can use water, methyl alcohol, ethanol or Virahol or its suitable mixture of basic solvent as be adjusted into alkaline pH, and can preferably use the water that is adjusted into pH9-12.
The extract obtained in this mode (elutant) can suitably be prepared as with preparation the appropriate form with raw material or medicament.For example, solution can be adjusted into and there is almost neutral pH, using as the preparation raw material, and can be adjusted into and there is expectation concentration by concentrated or dilution.In addition, about injection preparation, can add prepared by sodium-chlor etc. and ooze normal saline solution.In addition, but concentrate drying or freeze-drying solution, thus preparation can be used for the solid form of the raw material of tablet etc.
To the example of the patient's of needs treatments administration method comprise with pharmacology significant quantity oral administration with and other administration forms as subcutaneous, intramuscular and intravenously administration.Dosage can suitably be determined according to the type of the inflammation tissue extract of inoculating vaccinia virus.According to " Drugs in Japan, Ethical Drugs " (2978 pages), be obtained commercially the dosage ratified in preparation as prescription drugs basically oral administration with for 16NU every day, inject as every day 3.6 to 7.2NU.Yet dosage can be according to suitably increasing or reduce (NU: Neurotropin unit during disease type, severity, patient individual difference, administration method and administration etc.Neurotropin unit by use SART-stress mouse by the ED of the analgesic effect of improved Randall-Selitto method measurement 50value defines, and described stress mouse be the chronic stress animal that shows the low threshold of pain (pain threshold) than intact animal.1NU means ED 50value activity of 1mg analgesic composition in the Neurotropin preparation when the 100mg/kg preparation).
Hereinafter illustrate the inflammation tissue extract of producing the inoculation vaccinia virus method example and about the novel pharmacological activity of extract, that is, and to the pharmacology test result of the collagen protein in intervertebral disc cells and the synthetic promotion activity of proteoglycan.The present invention is not meant to the record content that is limited to following embodiment, wherein except contrary explanation, and all parts, per-cent and than all by weight, all temperature are ℃, and all pressure is normal atmosphere:
Embodiment 1
Cutaneous inoculation vaccinia virus by the healthy adult rabbit.By inflammation skin peeling crushing, and add phenol water to the skin of crushing.Then, depress filtering mixt adding, and with hydrochloric acid, gained filtrate is adjusted into to pH5, then heating 30 minutes under 90-100 ℃.After filtering deproteinated, with sodium hydroxide, filtrate is adjusted into to pH9, further under 90-100 ℃, heat 15 minutes, then filter.With hydrochloric acid, filtrate is adjusted into to about pH4.5, and adds 2% gac.Centrifugal after stirring the mixture 2 hours.Add water in the gac of collecting.With sodium hydroxide, mixture is adjusted into to pH10, stirs 1.5 hours under 60 ℃, then centrifuging, thus obtain supernatant liquor.Again add water in collected gac.With sodium hydroxide, mixture is adjusted into to pH11, under 60 ℃, stir after 1.5 hours centrifugal, thereby obtain supernatant liquor.By the combination of two kinds of supernatant liquors and with the hydrochloric acid neutralization, thereby obtain the extract of inflammation skin that the rabbit of vaccinia virus is arranged from inoculation.In following pharmacology test, extract is adjusted into to proper concn for use.
Embodiment 2
By the cutaneous inoculation vaccinia virus of healthy adult rabbit so that its infection.Subsequently, peel off and shred inflammation skin is aseptic, then add the glycerol liquor that is added with phenol.With homogenizer, mixture is milled to emulsus.Subsequently, emulsion is filtered.With hydrochloric acid, gained filtrate is adjusted into to slightly acidic (pH4.5-5.5), then under 100 ℃, heats and filter.With sodium hydroxide, filtrate is adjusted into to weakly alkaline (pH8.5-10.0), further heating under 100 ℃, then filter.With hydrochloric acid, filtrate is adjusted into to about pH4.5, and adds approximately 1.5% gac.Mixture is stirred after 1-5 hour and filters.To the gac of collecting by filtration, add water.With sodium hydroxide, mixture is adjusted into to pH9.4-10, stirs after 3-5 hour and filter.Use the hydrochloric acid neutralization filtrate.
Embodiment 3
Next, example about the pharmacology test result of the synthetic promotion activity to collagen protein in intervertebral disc cells and proteoglycan is shown, wherein the inflammation tissue extract (NTP) of the inoculation vaccinia virus of the present invention of acquisition in embodiment 1 is used as to substances.Hereinafter reach Fig. 1-4 and provide the effect of NTP to the cell proliferation in proteoglycan (PG) metabolism and collagen protein synthesis and intervertebral disc cells.
1. materials and methods
The ox nucleus pulposus (NP) and fibrous ring (AF) cell that from 14-18 month, large oxtail bone intervertebral disk, separate are encapsulated in (4E+6 cell/mL) in alginates, and at normal oxygen (21%O 2) or hypoxemia (5%O 2) in the DMEM/F12 that contains 10% foetal calf serum (FBS), cultivate 11 days under condition.Process these cultures 3 days with the NTP of three kinds of different concns (0.001NU/ml, 0.01NU/mL and 0.1NU/mL).In order to estimate the synthetic of PG and collagen protein, within the end 4 or 16 hours, use respectively 35s-sulfuric acid or 3h-proline(Pro) mark culture.As previously mentioned, collect bead and medium and by papain digestion [Masuda, the people such as K., J Orthop Res. after mark, 21,922-930 (2003), and Akeda, K, Spine, 31,959-966 (2006)], the independent incorporation (incorporation) of measuring the radioactivity precursor in medium and alginates bead.Use CellTiter96Aqueous One Solution Cell Proliferation Assay (Promega, Madison, WI) to estimate cell proliferation.Will be from all data normalizations of three experiments of the different batches of cell with the mean value of control group in each experiment.Use ANOVA to using the PSLD test as check (posthoc test) to estimate treatment effect afterwards.
2. result
1) cell proliferation
Time point at 7 and 14 days, oxygen concn and process the propagation do not affect NP and AF cell with NTP.
2) PG is synthetic
In the NP cell, at 5%O 2under cultivation, with the NTP processing, PG being synthesized significantly increases (under 0.01NU/ml ,+65%, p<0.05, under 0.1NU/ml ,+66%, p<0.05, Fig. 2).PG in the lower NP cell of normal oxygen cultivation synthesizes does not have notable difference.As shown in Figure 1, in the AF cell, under normal oxygen and hypoxia condition, NTP does not all change the synthetic level of PG.
3) collagen protein synthesis
As shown in Figures 3 and 4, process and make 5%O by NTP 2under NP cell and 21%O 2under NP and the collagen protein synthesis of AF cell increase (the highest stimulation under 0.1NU/ml: NP, 5%O in dosage dependence mode 2,+50%; NP, 21%O 2,+63%; AF, 21%O 2,+47%).
As implied above, this research illustrates NTP first can stimulate the extracellular matrix of intervertebral disc cells synthetic.What is interesting is, NTP is synthetic to the PG of NP cell acts on 5%O 2culture condition under than at 21%O 2culture condition under more obvious.Consider the interior hypoxia condition of body of NP, these results can show that the NP cell can respond NTP under the physiology correlated condition.Although at 5%O 2and 21%O 2under all observe the active effect of NTP to the collagen protein synthesis of NP cell, but the stimulation in AF is only at 21%O 2under condition, observe.Different in response to NTP from the synthetic increase of the PG of NP cell, the AF cell is at 21%O 2more respond NTP under condition.Because the oxygen tension in AF (oxygen tension) is significantly higher than NP, so 21%O 2lower AF cell also can react optimal culture condition to the larger response of NTP.
The chondrocyte is present in various cartilages, as joint cartilage, intervertebral disk, meniscus etc.The chondrocyte produces extracellular matrix along with cell fission around it, thereby cartilage is formed.Cartilage according to the content of [amorphous and fibre composition than being grouped into hyaline cartilage, elastic cartilage and fibrous cartilage.The main component of extracellular matrix is collagen protein and proteoglycan.Due to NTP the chondrocyte is promoted as the collagen protein in intervertebral disc cells and proteoglycan synthetic has active, so show that NTP has regeneration activity to the chondrocyte's of various cartilages extracellular matrix.
utilizability on industry
The present invention relates to inoculate the new medicine use of the inflammation tissue extract of vaccinia virus, particularly the chondrocyte is as the collagen protein in intervertebral disc cells, articular chondrocytes and meniscus cell and the synthetic promotion method of proteoglycan.Below provide the preferred embodiments of the invention:
1. the promotion method of the proteoglycan in a patient chondrocyte and/or collagen protein synthesis, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
2. the proteoglycan in a chondrocyte and/or the synthesis accelerant of collagen protein, it comprises the inflammation tissue extract of inoculating vaccinia virus.
3. the inflammation tissue extract of inoculation vaccinia virus is for promoting chondrocyte's the purposes of proteoglycan and/or collagen protein synthesis.
4. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, wherein used synthetic the promotions activity of proteoglycan in the chondrocyte who cultivates and/or collagen protein as index.
5. an inflammation tissue extract of inoculating vaccinia virus, it has the synthetic promotion of the proteoglycan in the chondrocyte and/or collagen protein active.
6. the renovation process of a chondrocyte extracellular matrix, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
7. the regenerator of a chondrocyte extracellular matrix, it comprises the inflammation tissue extract of inoculating vaccinia virus.
8. the purposes that the inflammation tissue extract of inoculation vaccinia virus is regenerated for the extracellular matrix that makes the chondrocyte.
9. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, wherein used chondrocyte's the regeneration activity of extracellular matrix as index.
10. an inflammation tissue extract of inoculating vaccinia virus, it has the regeneration activity of chondrocyte's extracellular matrix.
11. the synthetic promotion method of proteoglycan in an intervertebral disc cells (ox nucleus pulposus cell), it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
12. the promotion method of the collagen protein synthesis in an intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells), it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
13. the synthesis accelerant of the proteoglycan in an intervertebral disc cells (ox nucleus pulposus cell), it comprises the inflammation tissue extract of inoculating vaccinia virus.
14. the synthesis accelerant of the collagen protein in an intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells), it comprises the inflammation tissue extract of inoculating vaccinia virus.
15. the inflammation tissue extract of inoculation vaccinia virus is for promoting the synthetic purposes of intervertebral disc cells (ox nucleus pulposus cell) proteoglycan.
16. the inflammation tissue extract of inoculation vaccinia virus is for promoting the purposes of intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells) collagen protein synthesis.
17. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, wherein used the synthetic promotion of the proteoglycan in the intervertebral disc cells (ox nucleus pulposus cell) of cultivating active in index.
18. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, wherein used the synthetic promotion of the collagen protein in the intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells) of cultivating active in index.
19. an inflammation tissue extract of inoculating vaccinia virus, it has the synthetic promotion of the proteoglycan in intervertebral disc cells (ox nucleus pulposus cell) active.
20. an inflammation tissue extract of inoculating vaccinia virus, it has the synthetic promotion of the collagen protein in intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells) active.
21. the renovation process of the extracellular matrix of an intervertebral disc cells, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
22. the regenerator of the extracellular matrix of an intervertebral disc cells, it comprises the inflammation tissue extract of inoculating vaccinia virus.
23. the purposes that the inflammation tissue extract of inoculation vaccinia virus is regenerated for the extracellular matrix that makes intervertebral disc cells.
24. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, wherein used the regeneration activity of extracellular matrix of intervertebral disc cells as index.
25. an inflammation tissue extract of inoculating vaccinia virus, it has the regeneration activity of the extracellular matrix of intervertebral disc cells.
26. treatment or the prevention method of a sex change intervertebral disk (degenerative intervertebral discs), it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
27. the treatment of a sex change intervertebral disk or preventive, it comprises the inflammation tissue extract of inoculating vaccinia virus.
28. the purposes that the inflammation tissue extract of inoculation vaccinia virus is used for the treatment of or prevents the sex change intervertebral disk.
29. treatment or the prevention method of the pain caused by degeneration of intervertebral disc (degeneration), wherein make the extracellular matrix regeneration of intervertebral disc cells by the inflammation tissue extract of the inoculation of the patient's administration to this type for the treatment of of needs vaccinia virus.
30. the treatment of the pain caused by degeneration of intervertebral disc or prevention method, wherein the inflammation tissue extract by the patient's administration to this type for the treatment of of needs inoculation vaccinia virus promotes the synthetic of proteoglycan in intervertebral disc cells (ox nucleus pulposus cell).
31. the treatment of the pain caused by degeneration of intervertebral disc or prevention method, wherein the inflammation tissue extract by the patient's administration to this type for the treatment of of needs inoculation vaccinia virus promotes the synthetic of collagen protein in intervertebral disc cells (ox nucleus pulposus cell or annulus fibrosis cells).
32. the embodiment in secondary paragraph 1-31 is arbitrary as mentioned above, described inflammation is organized as the rabbit skin tissue.

Claims (26)

1. the proteoglycan in a chondrocyte and/or the promotion method of collagen protein synthesis, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
2. method according to claim 1, wherein said chondrocyte comprises intervertebral disc cells.
3. method according to claim 2, wherein said intervertebral disc cells comprises ox nucleus pulposus cell and/or annulus fibrosis cells.
4. method according to claim 2, wherein promote the synthetic of proteoglycan.
5. method according to claim 2, wherein promote the synthetic of collagen protein.
6. method according to claim 2, wherein said inflammation is organized as the rabbit skin tissue.
7. method according to claim 4, wherein said intervertebral disc cells comprises the ox nucleus pulposus cell.
8. method according to claim 4, wherein said intervertebral disc cells comprises annulus fibrosis cells.
9. method according to claim 5, wherein said intervertebral disc cells comprises the ox nucleus pulposus cell.
10. method according to claim 5, wherein said intervertebral disc cells comprises annulus fibrosis cells.
11. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, it comprises that the described extract of measurement is active in index to the synthetic promotion of the proteoglycan in the chondrocyte who cultivates and/or collagen protein.
12. method according to claim 11, wherein said chondrocyte comprises intervertebral disc cells.
13. method according to claim 12, wherein said intervertebral disc cells comprises ox nucleus pulposus cell and/or annulus fibrosis cells.
14. method according to claim 12, wherein promote the synthetic of proteoglycan.
15. method according to claim 12, wherein promote the synthetic of collagen protein.
16. method according to claim 12, wherein said inflammation is organized as the rabbit skin tissue.
17. the renovation process of a chondrocyte extracellular matrix, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
18. method according to claim 17, the extracellular matrix that wherein said chondrocyte's extracellular matrix is intervertebral disc cells.
19. judgement or an evaluation method of inoculating the inflammation tissue extract of vaccinia virus, it comprise measure described extract to the regeneration activity of chondrocyte's extracellular matrix as index.
20. method according to claim 19, the extracellular matrix that wherein said chondrocyte's extracellular matrix is intervertebral disc cells.
21. the methods for the treatment of of a sex change intervertebral disk, it comprises to the inflammation tissue extract of patient's administration inoculation vaccinia virus of this type for the treatment of of needs.
22. the methods for the treatment of of the pain caused by degeneration of intervertebral disc, it comprises that the inflammation tissue extract by the inoculation of the patient's administration to this type for the treatment of of needs vaccinia virus is regenerated the extracellular matrix of intervertebral disc cells.
23. the methods for the treatment of of the pain caused by degeneration of intervertebral disc, it comprises that inflammation tissue extract by the patient's administration to this type for the treatment of of needs inoculation vaccinia virus promotes the synthetic of proteoglycan in intervertebral disc cells.
24. the methods for the treatment of of the pain caused by degeneration of intervertebral disc, it comprises that inflammation tissue extract by the patient's administration to this type for the treatment of of needs inoculation vaccinia virus promotes the synthetic of collagen protein in intervertebral disc cells.
25. the treatment of a sex change intervertebral disk or preventive, it comprises the inflammation tissue extract of inoculating vaccinia virus.
26. the proteoglycan in an intervertebral disc cells and/or the synthesis accelerant of collagen protein, it comprises the inflammation tissue extract of inoculating vaccinia virus.
CN201180049567.2A 2010-10-14 2011-10-11 A method for promoting the synthesis of collagen and proteoglycan in chondrocytes Active CN103476923B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US39319010P 2010-10-14 2010-10-14
US61/393,190 2010-10-14
PCT/US2011/055757 WO2012051173A2 (en) 2010-10-14 2011-10-11 A method for promoting the synthesis of collagen and proteoglycan in chondrocytes

Publications (2)

Publication Number Publication Date
CN103476923A true CN103476923A (en) 2013-12-25
CN103476923B CN103476923B (en) 2015-04-29

Family

ID=45938923

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201180049567.2A Active CN103476923B (en) 2010-10-14 2011-10-11 A method for promoting the synthesis of collagen and proteoglycan in chondrocytes

Country Status (8)

Country Link
US (2) US20140255507A9 (en)
EP (1) EP2627343B1 (en)
JP (1) JP5937089B2 (en)
KR (1) KR101894754B1 (en)
CN (1) CN103476923B (en)
AU (1) AU2011316756B2 (en)
CA (1) CA2808927C (en)
WO (1) WO2012051173A2 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108601800A (en) * 2016-01-29 2018-09-28 国立大学法人千叶大学 The prevention of muscle damage or therapeutic agent
CN110179828A (en) * 2018-02-20 2019-08-30 学校法人东海大学 GalNAc transferase expression promotor containing Vaccination with vaccinia virus inflammatory tissue extract
CN111956669A (en) * 2020-09-07 2020-11-20 威世药业（如皋）有限公司 Preparation process of cowpox vaccine inflammation-causing rabbit fur extract injection

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI483729B (en) 2010-03-11 2015-05-11 Nippon Zoki Pharmaceutical Co Improving or therapeutic agent for chronic prostatitis, interstitial cystitis and/or urinary disturbance
CN103476923B (en) 2010-10-14 2015-04-29 日本脏器制药株式会社 A method for promoting the synthesis of collagen and proteoglycan in chondrocytes
CN115629217A (en) 2013-04-30 2023-01-20 日本脏器制药株式会社 Extract and preparation containing the same
US20170072321A1 (en) * 2013-05-22 2017-03-16 David S. Thompson Highly interactive fantasy sports interleaver
CA2987712A1 (en) * 2015-05-29 2016-12-08 Nippon Zoki Pharmaceutical Co., Ltd. Agent for promoting migration of pluripotent stem cells
JP6966763B2 (en) * 2015-06-30 2021-11-17 日本臓器製薬株式会社 Improvement or treatment of chondromalacia patella
JP7125732B2 (en) * 2016-09-09 2022-08-25 日本臓器製薬株式会社 Agent for improving or treating neurogenic bladder
JP7064197B2 (en) 2016-09-23 2022-05-10 国立大学法人大阪大学 Schwann cell differentiation promoter and peripheral nerve regeneration promoter
JP2019142853A (en) * 2018-02-20 2019-08-29 学校法人東海大学 Agent for promoting expression of n-acetylgalactosaminyltransferase containing extract from inflamed tissues inoculated with vaccinia virus

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100412188C (en) * 2003-02-21 2008-08-20 Uab研究基金会 Biologically active native biomatrix composition
CN100464782C (en) * 2002-10-31 2009-03-04 日本脏器制药株式会社 Remedy for fibromyalgia
US7556649B2 (en) * 2000-04-07 2009-07-07 Zimmer Orthobiologics, Inc. Methods and compositions for treating intervertebral disc degeneration
CN101528947A (en) * 2006-09-06 2009-09-09 日本脏器制药株式会社 Method for study, determination or evaluation by gene expression analysis

Family Cites Families (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS53101515A (en) 1977-02-17 1978-09-05 Nippon Zoki Pharmaceutical Co Medicine having anodyne * sedative and antiallergic activity and production thereof
JPS5587724A (en) 1978-12-27 1980-07-02 Nippon Zoki Pharmaceut Co Ltd Selective immune enhancer
JP2651674B2 (en) 1987-07-23 1997-09-10 日本臓器製薬 株式会社 New physiologically active substance and method for producing the same
US4985254A (en) 1987-11-06 1991-01-15 Nippon Zoki Pharmaceutical Co., Ltd. Method of treating ischemic diseases
JPH0825885B2 (en) 1988-04-15 1996-03-13 日本臓器製薬株式会社 Idiopathic thrombocytopenic purpura treatment
JP2539665B2 (en) 1988-06-20 1996-10-02 日本臓器製薬株式会社 Neurological drug
JP2539669B2 (en) 1988-07-15 1996-10-02 日本臓器製薬株式会社 Diabetic neuropathy treatment
JP2588109B2 (en) 1993-03-19 1997-03-05 日本臓器製薬株式会社 Painkillers
JP2594222B2 (en) 1993-09-28 1997-03-26 日本臓器製薬株式会社 New physiologically active substance-KF
JP3852786B2 (en) 1995-04-25 2006-12-06 日本臓器製薬株式会社 Bone atrophy improving agent
JP4033936B2 (en) 1997-01-08 2008-01-16 日本臓器製薬株式会社 Nitric oxide production inhibitor
JPH1180005A (en) 1997-09-12 1999-03-23 Nippon Zoki Pharmaceut Co Ltd Therapeutic agent for osteoporosis
JPH11139977A (en) 1997-11-07 1999-05-25 Nippon Zoki Pharmaceut Co Ltd Nef action suppressant
JP2000016942A (en) 1998-04-27 2000-01-18 Nippon Zoki Pharmaceut Co Ltd Therapeutic agent for ischemic disease
CN100425692C (en) 1998-06-02 2008-10-15 株式会社富吉摩托普拉泽兹 Method for preparing rabbitpox culturing tissue containing high-quality physiological activity
CN1055249C (en) 1998-07-15 2000-08-09 沈继平 Analgesia medicine and its mfg. method
JP2000143536A (en) 1998-11-13 2000-05-23 Nippon Zoki Pharmaceut Co Ltd Antihydropic agent
AU754989B2 (en) 1998-11-16 2002-11-28 Nippon Zoki Pharmaceutical Co., Ltd. A therapeutic agent for intractable vasculitis
JP2000336034A (en) 1999-03-19 2000-12-05 Nippon Zoki Pharmaceut Co Ltd Promoter for chemokine production
CN1272365A (en) 1999-04-15 2000-11-08 日本脏器制药株式会社 Physiological active material
TWI243055B (en) 2000-04-13 2005-11-11 Nippon Zoki Pharmaceutical Co Pharmaceutical composition for use in treatment of dermatitis
JP4711523B2 (en) 2001-02-13 2011-06-29 日本臓器製薬株式会社 Hypoalbuminemia improving agent
JP2004300146A (en) 2003-03-17 2004-10-28 Nippon Zoki Pharmaceut Co Ltd Preventive agent against infectious disease
KR101307999B1 (en) 2004-12-01 2013-09-12 니폰 조키 세야쿠 가부시키가이샤 Dried material and method for the manufacture thererof
US20060134646A1 (en) 2004-12-17 2006-06-22 Ansari Aftab A Method for treatment of HIV infection
TWI406664B (en) 2006-03-30 2013-09-01 Univ Kyoto Agent increasing the production of thioredoxin
JPWO2009028605A1 (en) 2007-08-31 2010-12-02 国立大学法人九州大学 Preventive or alleviating peripheral neuropathy with anticancer agents
TWI483729B (en) 2010-03-11 2015-05-11 Nippon Zoki Pharmaceutical Co Improving or therapeutic agent for chronic prostatitis, interstitial cystitis and/or urinary disturbance
CN103476923B (en) 2010-10-14 2015-04-29 日本脏器制药株式会社 A method for promoting the synthesis of collagen and proteoglycan in chondrocytes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7556649B2 (en) * 2000-04-07 2009-07-07 Zimmer Orthobiologics, Inc. Methods and compositions for treating intervertebral disc degeneration
CN100464782C (en) * 2002-10-31 2009-03-04 日本脏器制药株式会社 Remedy for fibromyalgia
CN100412188C (en) * 2003-02-21 2008-08-20 Uab研究基金会 Biologically active native biomatrix composition
CN101528947A (en) * 2006-09-06 2009-09-09 日本脏器制药株式会社 Method for study, determination or evaluation by gene expression analysis

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
NCBI: "Homo sapiens leukocyte cell derived chemotaxin 1 (LECT1), transcript variant 1, mRNA", 《GENBANK DATABASE》 *
夏欣一等: "COL 2A1基因突变与Ⅱ型胶原病", 《中国优生与遗传杂志》 *
席庆等: "CHONDROMODULIN-一族新的软骨特异性生长因子", 《口腔颌面修复学杂志》 *
樊碧发: "神经妥乐平在颈肩腰腿痛方面的应用", 《中国疼痛医学杂志》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108601800A (en) * 2016-01-29 2018-09-28 国立大学法人千叶大学 The prevention of muscle damage or therapeutic agent
CN108601800B (en) * 2016-01-29 2022-04-12 国立大学法人千叶大学 Prophylactic or therapeutic agent for muscle damage
CN110179828A (en) * 2018-02-20 2019-08-30 学校法人东海大学 GalNAc transferase expression promotor containing Vaccination with vaccinia virus inflammatory tissue extract
CN110179828B (en) * 2018-02-20 2023-10-31 学校法人东海大学 N-acetylgalactosamine transferase expression promoter containing vaccinia virus inoculated inflammatory tissue extract
CN111956669A (en) * 2020-09-07 2020-11-20 威世药业（如皋）有限公司 Preparation process of cowpox vaccine inflammation-causing rabbit fur extract injection

Also Published As

Publication number Publication date
CA2808927A1 (en) 2012-04-19
US10711292B2 (en) 2020-07-14
EP2627343A2 (en) 2013-08-21
US20160010135A1 (en) 2016-01-14
JP5937089B2 (en) 2016-06-22
CN103476923B (en) 2015-04-29
US20140255507A9 (en) 2014-09-11
EP2627343B1 (en) 2018-01-10
EP2627343A4 (en) 2014-08-06
WO2012051173A2 (en) 2012-04-19
AU2011316756A1 (en) 2013-03-14
KR20140020225A (en) 2014-02-18
KR101894754B1 (en) 2018-09-04
CA2808927C (en) 2018-10-30
JP2013544785A (en) 2013-12-19
US20130164383A1 (en) 2013-06-27
AU2011316756B2 (en) 2015-01-22
WO2012051173A3 (en) 2013-09-26

Similar Documents

Publication Publication Date Title
CN103476923B (en) A method for promoting the synthesis of collagen and proteoglycan in chondrocytes
Mukhamedshina et al. Mesenchymal stem cell therapy for spinal cord contusion: a comparative study on small and large animal models
CN101827617B (en) Compositions suitable for treatment of spinal disease, disorder or condition
CN102630157B (en) Injectable compositions for intra-articular use combining a viscosupplementation agent and a fibroblast growth medium
CN104105492B (en) The purposes of the fermentation preparation method of deer antler extract, thus obtained extract and the extract with anti-inflammatory activity
Bach et al. Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration
CN105748519A (en) Preparation method of adipose mesenchymal stem cell preparation for treating osteoarthritis
US20200254026A1 (en) Agent for promoting migration of pluripotent stem cells
Cao et al. Regulation of spinal neuroimmune responses by prolonged morphine treatment in a rat model of cancer induced bone pain
CN109963574A (en) It include composition of the fat stem cell source efflux body as active constituent for prevent or treat liver fibrosis
Xia et al. Continual deletion of spinal microglia reforms astrocyte scar favoring axonal regeneration
Qi et al. Radial extracorporeal shock wave therapy promotes osteochondral regeneration of knee joints in rabbits
CN103494808A (en) Application of melatonin in preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation
CN101969986A (en) Radical therapeutic agent for keloid and hypertrophic scar
Dufour et al. Direct perfusion improves redifferentiation of human chondrocytes in fibrin hydrogel with the deposition of cartilage pericellular matrix
CN103463128B (en) The preparation method and its usage of liquid in mammal medullary cell
Bhujel et al. An injectable engineered cartilage gel improves intervertebral disc repair in a rat nucleotomy model
CN103239380A (en) Application of traditional Chinese medicine, screening method and anti-ageing drug thereof
CN110179828B (en) N-acetylgalactosamine transferase expression promoter containing vaccinia virus inoculated inflammatory tissue extract
Shim et al. Enhancing cartilage regeneration through spheroid culture and hyaluronic acid microparticles: A promising approach for tissue engineering
CN110063972A (en) The application of Shexiang Baoxin Pills or Shexiang Baoxin Pills extract in preparation treatment neurodegenerative disease
Li et al. Homoplantaginin alleviates intervertebral disc degeneration by blocking the NF-κB/MAPK pathways via binding to TAK1
CN116549473A (en) Application of forsythin in preparation of medicine for treating intervertebral disc degeneration
Sharma The use and efficacy of biomimetic proteoglycans for cartilage repair and regeneration

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant